EZB-ICR Cell Line: A New Established and Characterized Oral Squamous Cell Carcinoma Cell Line From Tongue.

BACKGROUND: Tongue cancer is one of the most aggressive forms of oral squamous cell carcinoma which needs more investigations. Herein, we aimed to establish and characterize a tongue cancer cell line. METHODS: Tumor tissue was obtained from a 70-year-old woman with tongue cancer. The established cell line named as EZB-ICR and characterized for doubling time, expression of specific markers, HPV corporation and migration status using flow cytometry, immunofluorescence staining, multiplex PCR, and migration assay. RESULTS: EZB-ICR was negative for expression of mesenchymal specific markers, cytokeratin19, pan-cytokeratin, vimentin and EPCAM, but was positive for S100 and Nestin. No appearance of human papilloma virus DNA was seen. The doubling time of EZB-ICR was 31 hours and migration assay confirmed its metastatic nature. CONCLUSION: To the best of our knowledge, EZB-ICR is the first tongue human cancer cell line established in Iran, and its features make it appropriate for cancer-based in vitro studies and contribute to more studies on tongue cancer.
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Decreased levels of interleukin 27 in the serum of vitiligo patients.

BACKGROUND: Vitiligo is a common skin disorder in which melanocytes are destroyed by auto-reactive immune responses. The loss of melanocytes results in the appearance of depigmented areas in different parts of the body. Cytokines have remarkable roles in the pathogenesis of vitiligo, such as IL-1, IL-6, and TNF-α; interleukin 27 (IL-27) is a new member of the IL-6/IL-12 family, mainly released by activated antigen-presenting cells. IL-27 has been suggested to function as a pro-inflammatory as well as an anti-inflammatory cytokine. Altered concentrations of IL-27 have been shown in various auto-immune diseases such as multiple sclerosis, rheumatoid arthritis, and psoriasis. No studies have been conducted to determine the expression of this cytokine in vitiligo patients. OBJECTIVE: The objective of this study was to determine the serum concentration of IL-27 in vitiligo patients and compare it with normal individuals. METHODS: The serum concentration of IL-27 in 79 vitiligo patients was evaluated in comparison to 45 healthy controls using ELISA assay. RESULTS: Results showed decreased concentration of IL-27 in vitiligo patients as compared with healthy subjects (p=0.026). Furthermore, no correlation between IL-27 concentrations and disease parameters such as vitiligo severity and the extension of the depigmented area was observed. STUDY LIMITATION: A larger sample size would be more recommended for this study. CONCLUSION: The reduction in the serum levels of IL-27 in vitiligo patients compared to normal subjects suggested the possible anti-inflammatory role of this cytokine in vitiligo. Thus, IL-27 may be considered as a new target for the manipulation of the immune system in vitiligo patients.

Therapeutic effect of adipose-derived mesenchymal stem cells (ASCs) on radiation-induced skin damage in rats.

BACKGROUND: Radiation-induced skin injury remains a serious concern, which may limit the duration and dose of radiation treatment. The concept that stem cell injection may reduce tissue injury or assist its recovery after radiation has been recently argued. Herein, we examined the effect of adipose-derived mesenchymal stem cells (ASCs) on radiation-induced skin damage in rats. METHODS: This study is an experimental case control study. ASCs were isolated from peri uterine fat tissue of the rats. Then the rats received a 30 Gy single dose radiation to their buttocks skin using gamma radiation. Next day stem cells were transplanted subcutaneously in 16 rats as the case group. A group of 16 rats was considered as control group with radiation but no transplantation of stem cells. Then rats were examined and observed by macroscopic analysis and phenotypic scores during 4 weeks of follow up. RESULTS: The wound size in control group was significantly higher than case group in the second, third and fourth weeks of evaluation (P<0.05). There was no significant difference in skin lesion severity, pathological factors, and the onset of recovery signs between two groups (P>0.05). CONCLUSIONS: It seems that using ASCs alone has not profound effects on reducing radiation-induced cutaneous complications, while combination of these cells with growth factors may produce more promising results.

Comparison of Osteogenic and Chondrogenic Differentiation Ability of Buccal Fat Pad Derived Mesenchymal Stem Cells and Gingival Derived Cells.

STATEMENT OF THE PROBLEM: One major goal of tissue engineering and regenerative medicine is to find an appropriate source of mesenchymal stem cells (MSCs) with higher differentiation ability. PURPOSE: In this experimental study, the osteogenic and chondrogenic differentiation ability of buccal fat pad derived MSCs (BFP-MSCs) with gingival derived cells (GDCs) were compared. MATERIALS AND METHOD: BFP-MSCs and GDCs were cultured enzymatically and expanded. The expanded cells were analyzed for membrane-associated markers, using flow cytometry. Then the ability of these cells to differentiate into osteocyte and chondrocyte was assessed morphologically and by mRNA expression of collagen I (COLL), BGLA and bone morphogenetic protein 2 (BMP2) using qRT-PCR. RESULTS: Flow cytometry analysis showed that both BFP-MSCs and GDCs expressed the characteristic stem cell markers such as CD73, CD44, and CD90, whereas they did not express hematopoietic markers. Mineralized calcium deposition was observed apparently in BFP-MSCs cultured in osteogenic medium but GDCs showed fewer mineralized nodules. The mRNA expression levels of BGLA and BMP2 showed 7×105 and 733-fold more mRNA expression in BFP-MSCs treated with differentiation media compared to the control group. In chondrogenic differentiation, BFP-MSCs transformed from a spindle to a cuboidal shape while GDCs showed only a slight transformation. In addition, mRNA expression of COLL showed 282-fold higher expression in BFP-MSCs in comparison to the control group. Such significant difference in mRNA expression of BGLA, BMP2, and COLL was not observed in GDCs compared to their corresponding controls. CONCLUSION: Based on the present results, BFP yields a greater proportion of stem cells compared to gingiva. Therefore, this tissue can be introduced as an easily available source for the treatment of periodontal defects and other maxillofacial injuries.

Genistein Suppression of Matrix Metalloproteinase 2 (MMP-2) and Vascular Endothelial Growth Factor (VEGF) Expression in Mesenchymal Stem Cell Like Cells Isolated from High and Low Grade Gliomas

Objective: Brain tumors cause great mortality and morbidity worldwide, and success rates with surgical treatment remain very low. Several recent studies have focused on introduction of novel effective medical therapeutic approaches. Genistein is a member of the isoflavonoid family which has proved to exert anticancer effects. Here we assessed the effects of genistein on the expression of MMP-2 and VEGF in low and high grade gliomas in vitro. Materials and Methods: High and low grade glioma tumor tissue samples were obtained from a total of 16 patients, washed with PBS, cut into small pieces, digested with collagenase type I and cultured in DMEM containing 10% FBS. When cells reached passage 3, they were exposed to genistein and MMP-2 and VEGF gene transcripts were determined by quantitative real time PCR (qRT-PCR). Results: Expression of MMP-2 demonstrated 580-fold reduction in expression in low grade glioma cells post treatment with genistein compared to untreated cells (P value= 0.05). In cells derived from high grade lesions, expression of MMP-2 was 2-fold lower than in controls (P value> 0.05). Genistein caused a 4.7-fold reduction in VEGF transcript in high grade glioma cells (P value> 0.05) but no effects were evident in low grade glioma cells. Conclusion. Based on the data of the present study, low grade glioma cells appear much more sensitive to genistein and this isoflavone might offer an appropriate therapeutic intervention in these patients. Further investigation of this possibility is clearly warranted.

Decreased levels of interleukin 27 in the serum of vitiligo patients

Abstract Background: Vitiligo is a common skin disorder in which melanocytes are destroyed by auto-reactive immune responses. The loss of melanocytes results in the appearance of depigmented areas in different parts of the body. Cytokines have remarkable roles in the pathogenesis of vitiligo, such as IL-1, IL-6, and TNF-α; interleukin 27 (IL-27) is a new member of the IL-6/IL-12 family, mainly released by activated antigen-presenting cells. IL-27 has been suggested to function as a pro-inflammatory as well as an anti-inflammatory cytokine. Altered concentrations of IL-27 have been shown in various auto-immune diseases such as multiple sclerosis, rheumatoid arthritis, and psoriasis. No studies have been conducted to determine the expression of this cytokine in vitiligo patients. Objective: The objective of this study was to determine the serum concentration of IL-27 in vitiligo patients and compare it with normal individuals. Methods: The serum concentration of IL-27 in 79 vitiligo patients was evaluated in comparison to 45 healthy controls using ELISA assay. Results: Results showed decreased concentration of IL-27 in vitiligo patients as compared with healthy subjects (p = 0.026). Furthermore, no correlation between IL-27 concentrations and disease parameters such as vitiligo severity and the extension of the depigmented area was observed. Study limitation: A larger sample size would be more recommended for this study. Conclusion: The reduction in the serum levels of IL-27 in vitiligo patients compared to normal subjects suggested the possible anti-inflammatory role of this cytokine in vitiligo. Thus, IL-27 may be considered as a new target for the manipulation of the immune system in vitiligo patients.

Expression of chemokines and chemokine receptors in brain tumor tissue derived cells.

Chemokine and chemokine receptor expression by tumor cells contributes to tumor growth and angiogenesis and thus these factors may be considered as tumor markers. Here we aimed to characterize cells directly extracted from glioma, meningioma, and secondary brain tumors as well as non-tumoral cells in vitro. Cells were isolated from brain tissues using 0.2% collagenase and characterized by flow cytometry. Expression of SDF-1, CXCR4, CXCR7, RANTES, CCR5, MCP-1 and IP-10 was defined using flow cytometry and qRT-PCR methods. Brain tissue isolated cells were observed as spindle-shaped cell populations. No significant differences were observed for expression of SDF-1, CXCR4, CXCR7, RANTES, CCR5, and IP-10 transcripts. However, the expression of CXCR4 was approximately 13-fold and 110-fold higher than its counterpart, CXCR7, in meningioma and glioma cells, respectively. CXCR7 was not detectable in secondary tumors but CXCR4 was expressed. In non tumoral cells, CXCR7 had 1.3-fold higher mRNA expression than CXCR4. Flow cytometry analyses of RANTES, MCP- 1, IP-10, CCR5 and CXCR4 expression showed no significant difference between low and high grade gliomas. Differential expression of CXCR4 and CXCR7 in brain tumors derived cells compared to non-tumoral samples may have crucial impacts on therapeutic interventions targeting the SDF-1/CXCR4/CXCR7 axis.

Scaffold-free adipose-derived stem cells (ASCs) improve experimentally induced osteoarthritis in rabbits.

BACKGROUND: Osteoarthritis (OA) is the most common form of arthritis seen clinically. Current treatments for OA are limited to decreasing associated pain, maintaining or improving joint function, and minimizing disability. However, these treatments have no effect on the regeneration of hyaline cartilage. Since mesenchymal stem cells (MSCs) have been described as promising cell sources for cartilage repair, the present study was designed to examine whether intra-articular injection of scaffold-free adipose-derived stem cells (ASCs) obtained from subcutaneous adipose tissue could restore the matrix of arthritic knee joints in mature animals. METHODS: OA was induced in adult white New Zealand rabbits by unilateral anterior cruciate ligament transection (ACLT); the contralateral knee was considered the sham-operated group. At 12 weeks following surgery, the ASCs treated group was injected intra-articularly with a single dose of 1 × 10(6) cells suspended in 1 mL of medium. The control group received 1 mL of medium without cells and the sham-operated group received no treatment. All rabbits were sacrificed at 16 and 20 weeks after surgery. OA progression was evaluated radiologically, grossly, and histologically using hematoxylin and eosin, Safranin-O, and toluidine blue staining. RESULTS: At 12 weeks after surgery all knees subjected to ACLT showed radiological signs of OA. The findings showed significant differences in the quality of cartilage between ASCs-injected group compared to control group, particularly at 20 weeks after surgery. CONCLUSION: This study suggests that ASCs obtained from subcutaneous adipose tissue could be a viable approach for treating OA.